JP2679737B2 - L-glutamic acid derivative - Google Patents

Abstract

Pyrimidin-5-yl derivatives of L-glutamic acid are anti-neoplastic agents. A typical embodiment is N-(4-[4-(2,4-diamino-6-hydroxypyrimidin-5-yl)-butyl]-benzoyl)-L-glutamic acid.

Description

The present invention relates to an L-glutamic acid derivative which is an antineoplastic agent, a method for producing the same and uses thereof, and an intermediate useful in the production thereof.

BACKGROUND OF THE INVENTION The antifolates aminopterin and amethopterin (also known as 10-methylaminopterin or methotrexate) are antitumor agents. These compounds inhibit the oxygen conversion involved in metabolic derivatives of folic acid.
For example, amethopterin inhibits dihydrofolate reductase, which is a necessary enzyme for the regeneration of tetrahydrofolate from dihydrofolate formed during the conversion of 2-deoxyuridylic acid to thymidylate by the thymidylate synthetase enzyme.

Folic acid and other derivatives of aminopterin have been synthesized and tested as antimetabolites. Among these are compounds in which a methylene or methylidene group occupies a position in the molecule that is normally occupied by an imino or nitrilo group, respectively. These derivatives have varying degrees of antimetabolite activity. 10-Deazaaminopterin is highly active [Sirotaku et al., Cancer Treatment Report, 1978, 62, 1047 (Sirotak et al., Cancer
Treatment Report, 1978, 62, 1047)], 5-deazaaminopterin has an activity similar to that of amethopterin [Taylor et al., Journal of Organic Chemistry, 1983, Vol. 48, Vol. Page 4852 (Taylor
et al., Journal of Organic Cemistry, 1983,48,485
2)]. 8,10-Dideazaaminopterin was reported to be active (US Pat. No. 4,460,591), 5,8,1.
0-Trideazaaminopterin is active against mouse L1210 leukemia [Yan et al., Journal of Heterocyclic Chemistry, 1979, 16: 541 (Yan et al., Journal of Heterocyclic Cemistry. , 197
9,16,541)]. On the other hand, 10-deazafolate does not show significant activity [Strack et al., Journal of Medicinal Chemistry, 1971, Vol. 14, p. 693 (Struck et al.
al., Journal of Medicinal Cemistry, 1971,14,69
3)], 5-deazafolate is only weakly cytotoxic. 8,1
0-Dideazafolate is only slightly effective as a dihydrofolate reductase inhibitor [De Gra (De Gra
w) et al., "Chemistry and Biology of Pteridines," Elsevier, 1979, p. 229], 5,8,10-Trideazafolate also shows little activity against mouse L1210 leukemia [Otis (Otis Oatis) et al., Journal of
Medicinal Chemistry, 1977, Volume 20, 139
page〕. 5,10-Dideazaaminopterin and 5,10-dideaza-5,6,7-8-tetrahydroaminopuritene, and the corresponding 5,10-dideazafolate derivatives are described in Taylor et al., Journal of Medicinal Chemistry, 28th
Vol. 7, No. 914, 1985.

DISCLOSURE OF THE INVENTION The present invention provides glutamic acid derivatives of the formula: [Wherein each of X and Y independently of the other is hydroxy or amino; Z is unsubstituted or substituted with 1 to 4 chlorine or fluorine atoms, 1,4-phenylene; cyclohexa-1, 4-diyl; or a linear or branched alkylene group having 2 to 5 carbon atoms; n has a value of 2 to 6; the carbon atom arrangement indicated by * is L] and its pharmaceutically acceptable Related to salt.

The present invention also relates to methods of making such compounds, intermediates useful in their manufacture, and methods and compositions for the use of such compounds to combat tumor growth.

Modes for Carrying Out the Invention The compounds of formula I exist in tautomeric equilibrium with the corresponding 4-oxo and 4-imino compounds: For convenience, throughout this specification 3,4-dehydro-4-
The hydroxy and 3,4-dehydro-4-amino forms have been described and the corresponding nomenclature has been used, but in each case they include the tautomeric 4 (3H) -oxo and imino forms. It should be understood.

The symbol n can have values from 2 to 6 and therefore ethylene, trimethylene, 1,2-propylene, 2,3-propylene, tetramethylene, 1,2-butylene, 2,3-butylene, pentamethylene, 1 2,2-Pentylene, 2,3-Pentylene, 1,3-Pentylene, Hexamethyene, 1,2-Hexylene, 2,3-Hexylene, 2,4-Hexylene and other divalent alkylene groups having 2 to 6 carbon atoms. Have. Preferably,
n has a value of 3-5. Further, when n is 4 or 5, the two monovalent carbon atoms are separated by two carbon atoms, ie, tetramethylene, 1,4-pentylene,
2,5-Pentylene and pentamethylene are preferred. 1,4-
It is understood that certain branched alkylene groups such as pentylene may be oriented in either of two ways such that the branched carbon atom is adjacent to the pyrimidin-5-yl group or adjacent to Z. Will

The radical Z is a hydrocarbon radical which may be unsubstituted or substituted with 1 to 4 chlorine or fluorine atoms, such as 1,4-phenylene, for example -chloro-1,4-phenylene, 2-fluoro-1,4. -Phenylene, 3-chloro-1,4-phenylene, 3
-Fluoro-1,4-phenylene, 2,6-dichloro-1,4-
Phenylene, 2,6-difluoro-1,4-phenylene, 3,5
-Dichloro-1,4-phenylene, 3,5-difluoro-1,4
-Phenylene, 2,3,5,6-terolafluoro-1,4-phenylene and the like. Z is cyclopenta-1,4-diyl or an alkylene group having 2 to 5 carbon atoms, such as ethano, trimethylene, 1,3-butylene, 2,4-butylene, 1,3-pentylene, 2,4-pentylene or 3 It may also be 5,5-pentylene.

Each of X and Y is hydroxy or amino. Preferably Y is amino, most preferably Y is amino and X is hydroxy.

In the present invention, pharmaceutically acceptable alkali metal, alkaline earth metal, non-toxic metal, ammonium and substituted ammonium salts such as sodium, potassium, lithium,
Calcium, magnesium, aluminum, zinc, ammonium, trimethylammonium, triethylammonium, triethanolammonium, pyridinium,
Included are salts such as substituted pyridiniums and the like.

The compound may be prepared in an initial process by hydrolysis of a 2-aminopyrimidin-5-yl-L-glutamic acid derivative of the formula: In the above formula, each of R ² and R ³ is a carboxylic acid protecting group; and X, Y, Z and n are as defined above.

The protecting groups contained in R ² and R ³ and their removal reactions are described, for example, in “Protecting Groups in Organic Chemistry”, Plenum Press,
London and New York, 1973 (“Protective Grou
ps in Organic Chemistry ", Plenum Press, London and N
ew York (1973)]; Green “Protecting groups in organic synthesis” Willy, New York, 1981 [Greene, “Protec
tive Groups in Organic synthesis ", Wiley, New York
(1981); Peptides, Volume I, Schrader and Lubke, Academic Press, London and New York, 1965 ["The Peptides", Vol. I. Schrder and Lub.
ke, Academic Press, London and New York (1965)];
"Methods of Organic Chemistry", Hoven-Weil, 4th edition, 15th.
Volume I, Georg Team Ferluk, Stadtgart, 1974 ["Methoden der Organische Chemie", Hou
ben−Wey1,4th Edition, Vol.15 / I, Georg Thieme Verla
g, Stuttgart (1974)]. Carboxylic acid protecting groups are conceptually derived from lower alkanols having 1 to 6 carbon atoms, including those branched at the 1-position and those substituted with one or more aromatic groups such as phenyl or substituted with halo or alkoxy. Esters such as methyl, ethyl, t-butyl, benzyl, 4-nitrobenzyl, diphenylmethyl, methoxymethyl and the like. Silyl esters such as trimethylsilyl can also be used.

Hydrolysis may be carried out in the presence of a water-miscible organic solvent such as methanol, ethanol, tetrahydrofuran, dimethylformamide or the like or an acid such as trifluoroacetic acid or an aqueous acid such as an aqueous alkali metal hydroxide. It is carried out at room temperature using a base. When a base is used, the product is initially formed as the dicationic glutamate salt, but can be easily precipitated by adjusting the pH, for example via acidification with acetic acid. The product obtained is usually a crystalline or microcrystalline solid.

Compounds of formula II are prepared by conventional condensation techniques for peptide bond formation, such as formation of mixed anhydrides, treatment with DCC or activation of the carboxylic acid group by use of diphenylchlorophosphonate. : The protected glutamic acid derivative of the following formula: It can be manufactured by coupling with.

Formation of the intermediate of formula III is accomplished by using the guanidine free base to form the α-cyanodicarboxylate of the formula: [Wherein R ⁴ R ⁴ ′ is the same or different alkoxy group having 1 to 6 carbon atoms; n and Z are as defined above].

The intermediate of formula V is obtained by converting an alkyl cyanoacetate of the following formula: R ⁴ CO—CH ₂ —CN (VI) into a mesityl ester of the following formula: CH ₃ SO ₂ O in the presence of a strong base such as sodium hydride. - it is prepared by condensing a _{(C n H 2n) -Z-} COR 4 '(VII).

Meshichiruesuru intermediates of formula VII, hydroxy acid esters of the formula: HO- but _{(C n H 2n) -Z-} COR 4 ' can be obtained from (VIII), which is more fully at or below are known It can be produced by a known method as exemplified in.

Representative compounds of the present invention are as follows: N- [4- [3- (2,4-diamino-6-hydroxypyrimidin-5-yl) propyl] benzoyl] -L-glutamic acid; N- [4 -[4- (2,4-Diamino-6-hydroxypyrimidin-5-yl) butyl] benzoyl] -L-glutamic acid; N- [4- [2-chloro-4- (2,4-diamino-6-
Hydroxypyrimidin-5-yl) butyl] benzoyl] -L-glutamic acid; N- [4- [2-puroro-4- (2,4-diamino-6)
-Hydroxypyrimidin-5-yl) butyl] benzoyl] -L-glutamic acid; N- [4- [3-chloro-4- (2,4-diamino-6-
Hydroxypyrimidin-5-yl) butyl] benzoyl] -L-glutamic acid; N- [4- [3-fluoro-4- (2,4-diamino-6)
-Hydroxypyrimidin-5-yl) butyl] benzoyl] -L-glutamic acid; N- [4- [2,6-difluoro-4- (2,4-diamino-
6-hydroxypyrimidin-5-yl) butyl] benzoyl] -L-glutamic acid; N- [4- [3,5-difluoro-4- (2,4-diamino-
6-Hydroxypyrimidin-5-yl) butyl] benzoyl] -L-glutamic acid; N- [4- [5- (2,4-diamino-6-hydroxypyrimidin-5-yl) pentyl] benzoyl] -L-glutamic acid N- [4- [3- (2,4,6-triaminopyrimidine-5
-Yl) propyl] benzoyl] -L-glutamic acid; N- [4- [4- (2,4,6-triaminopyrimidine-5
-Yl) butyl] benzoyl] -L-glutamic acid; N- [4- [5- (2,4,6-triaminopyrimidine-5
-Yl) pentyl] benzoyl] -L-glutamic acid; N- [4- [4- (2,4-diamino-6-hydroxypyrimidin-5-yl) propyl] cyclohexylcarbonyl] -L-glutamic acid; N- [8- (2,4-Diamino-6-hydroxypyrimidin-5-yl) octanoyl] -L-glutamic acid; N- [5- (2,4-diamino-6-hydroxypyrimidin-5-yl) pentanoi]- L-glutamic acid; N- [6- (2,4-diamino-6-hydroxypyrimidin-5-yl) hexanoyl] -L-glutamic acid; N- [7- (2,4-diamino-6-hydroxypyrimidine-5 -Yl) heptanoyl] -L-glutamic acid; N- [9- (2,4-diamino-6-hydroxypyrimidin-5-yl) nonanoyl] -L-glutamic acid; N- [10- (2 , 4-Diamino-6-hydroxypyrimidin-5-yl) decanoyl] -L-glutamic acid; N- [11- (2,4-diamino-6-hydroxypyrimidin-5-yl) undecanoyl] -L-glutamic acid;
And N- [4- [4- (2-amino-4,6-dihydroxypyrimidin-5-yl) butyl) benzoyl] -L-glutamic acid.

The compounds of formula I contain a chiral center at the L-glutamic acid moiety in the molecule. - (C _n H _2n) - the presence of or even one or more chiral centers in one or both of Z results diastereomers. These diastereomers are
Each may be mechanically separated, such as by chromatography, so that each is substantially free of the other, ie, having an optical purity of greater than 95%. On the other hand, mixtures of diastereoisomeric compounds of formula I are treated with a chiral acid which can be manipulated to form salts with them. The resulting diastereoisomeric salt is then separated by one or more fractional crystals, after which the free base of the cationic portion of at least one separated salt is liberated by treatment with base and removal of the protecting groups. . The release of the cation of the salt can be carried out as a separate step before or after the removal of the protecting group, or simultaneously with the removal of such a group under basic conditions, ie if it is susceptible to basic hydrolysis.

Suitable chiral acids include individual enantiomers such as 10-camphorsulfonic acid, camphoric acid, α-bromocamphoric acid, methoxyacetic acid, tartaric acid, diacetyltartaric acid, malic acid, pyrrolidone-5-carboxylic acid.

The compounds of the present invention are effective with respect to one or more enzymes which utilize folic acid as a substrate and especially metabolic derivatives of folic acid.

The compounds of formula I are associated with choriocarcinoma, leukemia, adenocarcinoma of the female breast,
It can be used alone or in combination to treat tumors including epidermal cancers of the head and neck, squamous shoulder epithelial or small cell lung cancer and various lymphosarcoma. For example, in a typical model, N- [4- (2,4-diamino-
6-hydroxypyrimidine-5-butyl) benzoyl]
-L-Glutamic acid showed an IC ₅₀ value of 0.0632 μg / ml against the CCRF-CEM cell line (human T cell-derived leukemia). The compounds can also be used to treat dentivosis, psoriasis and arthritis.

The compounds are administered either alone or preferably parenterally to a mammal having a tumor and in need of treatment, either alone or in combination with other anti-tumor agents, steroids and the like. Parenteral routes of administration include intramuscular and intrathecal (intratheca
l), intravenous or intraarterial. In general, the compounds are often administered in a manner similar to methotrexate, but because of the different mode of action, they can be administered at higher doses than normally used for methotrexate. The dosage regimen will depend on the particular tumor, the condition of the patient and the responsiveness, with the usual dosage being 5
Dosage is from about 10 to about 100 mg / day for up to 10 days or is repeated cyclically, for example 250-500 mg once daily every 14th day. Oral dosage forms include tablets and capsules containing 1-10 mg of drug per unit dose. Isotonic solutions containing 20-100 mg / ml can be used for parenteral administration.

The invention will be further illustrated by the following examples.

Example 1 65.07 g (1.0 eq) methyl 4- (4-hydroxybutyl) benzoate and 33.20 g triethylamine in 350 ml anhydrous ethyl ether stirred under nitrogen and cooled to 0 ° C.
37.58 g of mesityl chloride in the form of a drop wise addition to a mixture of
(1.05 eq) is added. A solid forms, but the reaction mixture is allowed to come slowly to room temperature and stirred under nitrogen for 4 hours. Then 400 ml of water was added, the organic layer was separated, washed with water, dried over magnesium sulfate and concentrated to give 4-mp of mp 52-53 ° C.
Obtain methyl (4-mesitylbutyl) benzoate; 1R (KB
r) ν _max 3010,2930,2850,1700,1603,1432,1334,1323,11
65,1104,937,822 and 700 cm ^-1 ; ¹ H NMR (CDCl ₃ ) δ 7.92 (d, J = 8.2 Hz, 2H, Ar), 7.22
(DJ = 8.2 Hz, 2H, Ar), 4.18-4.22 (m, 2H, CH ₂ OMs), 3.
86 (s, 3H, OCH ₃ ), 2.96 (s, 3H, SO ₂ CH ₃ ), 2.65-2.68 (m.
2H, benzyl), 1.72-1.74 (m, 4H, 2 ° aliphatic). Calculated value (C ₁₃ H ₁₈ SO ₅ ): C 54.53; H 6.34; S 11.20. Found: C 54.79; H 6.46; S 11.17. Methyl 4- (3-mesitylpropyl) benzoate, 4-
Methyl (5-mesitylpentyl) benzoate and methyl 10-mesityldecanoate can be prepared in a similar manner.

Example 2 A mixture of 10.30 g (1.1 eq) of 80% sodium hydride in mineral oil is washed twice with anhydrous tetrahydrofuran (to remove the mineral oil), after which 400 ml of tetrahydrofuran are added. It is cooled to 0 ° C. and a solution of 35.29 g (1.0 eq) ethyl cyanoacetate in anhydrous tetrahydrofuran is added dropwise under nitrogen. The mixture is stirred vigorously at ambient temperature until hydrogen evolution ceases, then 4- (4-mesitylbutyl).
89.33 g (1.0 eq.) Methyl benzoate in a non-aqueous solution is added dropwise. The mixture is stirred at ambient temperature for 12 hours, then at reflux temperature for 6 hours and then cooled. The solvent is removed under reduced pressure and diethyl ether is added. Separating the organic phase,
Wash with water and brine, dry over magnesium sulfate, filter, and concentrate. The residue is chromatographed with 1: 4 ethyl acetate: hexane, the homogenous fractions are combined, concentrated and
Methyl 4- (5-carbetoxy-5-cyanopentyl) benzoate is obtained as a liquid; 1R (film) ν _max 2915,2860,2255,1730,1609,1571,1
432,1413,1368,1320,1178,1105,1019,962,853,759 and 6
02cm ^-1 ; ¹ H NMR (CDCl ₃ ) δ7.95 (d, J = 8.1 Hz, 2H, Ar),
7.24 (d, J = 8.1 Hz, 2H, Ar), 4.25 (q, J = 7.2 Hz, 2H, C H
₂ CH ₃ ), 3.90 (s, 3H, CO ₂ CH ₃ ), 3.45 (t, J = 6.8 Hz, 1H, CH
CN), 2.69 (t, J = 7.5 Hz, 2H, engine), 1.93−2.00
( M , 2H, C H ₂ CH), 1.54-1.72 (m, 4H, 2 ° aliphatic), 1.30
(T, J = 7.3 Hz, 3H, CH ₂ C H ₃ ). Analytical calculation (C ₁₇ H ₂₁ NO ₄ ): C 67.31; H 6.98; N 4.62. Found: C 67.10; H 6.96; N 4.33. Methyl 4- (4-carbetoxy-4-cyanobutyl) benzoate, 4- Methyl (6-carbetoxy-6-cyanohexyl) benzoate and methyl 11-carbetoxy-11-cyanoundecanoate can also be prepared in a similar manner.

Example 3 A mixture of 0.71 g (1.05 eq) of guanidine hydrochloride, 0.40 g (1.05 eq) of sodium methoxide and 20 ml of anhydrous methanol is stirred under nitrogen for 30 minutes. The solid formed is filtered off and the filtrate is concentrated under reduced pressure. 2.15 g of methyl 4- (5-carbetoxy-5-cyanopentyl) benzoate in 20 ml of anhydrous dimethylformamide was added to the residue (guanidine free base).
Add (1.0 equivalent). The mixture is stirred under nitrogen with gentle heating for 12 hours, cooled and then mixed with very dilute sulfuric acid. The solid was collected, washed with water and then with diethyl ether, dried and mp203-205 ° C 4- [4-
Methyl (2,4-diamino-6-hydroxypyrimidin-5-yl) butyl] benzoate is obtained; 1R (KBr) ν _max 3495,3380,2925,2850,1685,1600,1490,1
430,1359,1282,1174,1111,1015 and 754 cm ^-1 ; ¹ H NMR (d ⁶
DMSO) δ9.74 (bs, 1H, (3) -NH), 7.83 (d, J = 8.1 H
z, 2H, Ar), 7.31 (d, J = 8.1 Hz, 2H, Ar), 5.87 (bs, 2H,
(2) -NM ₂ ), 5.60 (bs, 2H, (6) -NH ₂ ), 3.80 (s, 3
H, CH ₃ ), 2.62 (t, J = 7.6 Hz, 2H, (5) −CH ₂ ), 2.15
(T, J = 7.2 Hz, 2H, benzyl), 1.51-1.55 (m, 2H, 2 ° aliphatic), 1.26-1.28 (m, 2H, 2 ° aliphatic). Analytical calculation (C ₁₆ H ₂₀ N ₄ O ₃ ): C 60.75; H 6.37; N 17.71. Found: C 61.05; H 6.45; N 17.46. 4- [3- (2,4-diamino-6-hydroxy Methyl pyrimidin-5-yl) propyl] benzoate, 4- [5
-(2,4-Diamino-6-hydroxypyrimidine-5-
Methyl) pentyl] benzoate and methyl 10- (2,4-diamino-6-hydroxypyrimidin-5-yl) decanoate can be prepared in a similar manner.

Example 4 Methyl 4- [4- (2,4-diamino-6-hydroxypyrimidin-5-yl) butyl] benzoate 0.91 g and 1N
While gently heating a mixture of 40 ml of sodium hydroxide
Stir for 18 hours, then filter. Acidification with glacial acetic acid gives a solid which is collected by centrifugation with water. By concentrating under reduced pressure, 4- [4-
(2,4-Diamino-6-hydroxypyrimidin-5-yl) butyl] benzoic acid is obtained; IR (KBr) ν _max 3460,3345,3140,2920,2845,1575,1435,1
368,1247,1170 and 744 cm ^-1 ; ¹ H NMR (d ⁶ DMSO) δ 9.78 (b
s, 1H, (3) -NH), 7.80 (d, J = 8.0 Hz, 2H, Ar), 7.26
(D, J = 8.0 Hz, 2H, Ar), 5.88 (bs, 2H, (2) −NH ₂ ), 5.
59 (bs, 2H, (6 ) -NH 2), 2,61 (t, J = 7.4 Hz, 2H,
(5) -CH ₂ ), 2.15 (t, J = 7.2 Hz, 2H, benzyl), 1.51
-1.56 (m, 2H, 2 ° aliphatic), 1.27-1.31 (m, 2H, 2 ° aliphatic). Analytical calculation (C ₁₅ H ₁₈ N ₄ O ₃ ): C 59.59; H 6.00; N 18.53. Found: C 59.79; H 5.77; N 18.30. 4- [3- (2,4-diamino-6-hydroxy) Pyrimidin-5-yl) propyl] benzoic acid, 4- [5- (2,
4-diamino-6-hydroxypyrimidin-5-yl)
Pentyl] benzoic acid and 10- (2,4-diamino-6-hydroxypyrimidin-5-yl) decanoic acid can also be prepared in a similar manner.

Example 5 0.47 g (1.0 equivalent) of 4- [4- (2,4-diamino-6-hydroxypyrimidin-5-yl) butyl] benzoic acid,
Phenyl N-phenylphosphoramide chloridate 0.62
g (1.5 equivalents), N-methylmorpholine 0.79 g (5.0 equivalents)
And a mixture of 50 ml of anhydrous N-methylpyrrolidone are stirred under nitrogen at ambient temperature for 1 hour. Then, L-glutamic acid diethyl hydrochloride 0.75 g (2.0 eq) was added and stirring was done under nitrogen.
Continue for 24 hours. The solvent is removed by vacuum distillation and chloroform is added to the residue. The chloroform solution is washed with water, dried over magnesium sulfate, filtered and concentrated under reduced pressure. The residue was chromatographed with 1: 9 methanol: chloroform, N- [4- [4- (4- (2,4-diamino-6-hydroxypyrimidin-5-yl) butyl] at mp 75-76 ° C.
Benzoyl] -diethyl L-glutamate is obtained; IR (K
Br) ν _max 3320,2920,2915,1845,1720,1580,1529,1427,1
368,1326,1192,1093,1021,849 and 750 cm ^-1 ; ¹ H NMR (CDC
l ₃ ) δ 11.58 (bs, 1H, (3) -NH), 7.68 (d, J = 7.9 Hz,
2H, Ar), 7.33 (d, J = 7.9 Hz, 1H, NH), 7.17 (d, J = 7.9 H
z, 2H, Ar), 5.54 (bs, 2H, (2) -NH 2), 4.76-4.82 (m,
1H, CH), 4.71 (bs , 2H, (6) -NH 2), 4.23 (q, J = 7.11
Hz, 2H, CO ₂ CH ₂ ), 4.12 (q, J = 6.5 Hz, 2H, CO ₂ CH ₂ ), 2.15
-2.68 (m, 8H, 2 ° aliphatic), 1.57-1.66 (m, 2H, 2 ° aliphatic), 1.35-1.45 (m, 2H, 2 ° aliphatic), 1.30 (t, J = 7.1H
z, 3H, CH ₃ ), 1.22 (t, J = 7.1 Hz, 3H, CH ₃ ); C ₂₄ H ₃₃ N ₅ O
HRMS calculated for ₆ (M +): 487.2430; found: 487.24
15; m / e Other ions at 348,313,285,195,167,139,97 and 84. Calculated value (C ₂₄ H ₃₃ N ₅ O ₆ ): C 59.12; H 6.82; N 14.36. Found value: C 59.08; H 6.79; N 14.11. N- [4- [3- (2,4-diamino- 6-Hydroxypyrimidin-5-yl) propyl] benzoyl] -L-
Diethyl glutamate, N- [4- [5- (2,4-diamino-6-hydroxypyrimidin-5-yl) pentyl] benzoyl] -L-glutamate diethyl and N-
[10- (2,4-diamino-6-hydroxypyrimidine-
5-yl) decanoyl] -diethyl L-glutamate is similarly prepared.

Example 6 A mixture of 0.50 g of N- [4- (2,4-diamino-6-hydroxypyrimidin-5-yl) butyl] benzoyl] -diethyl L-glutamate and 40 ml of 1N sodium hydroxide is stirred at room temperature for 72 hours. The mixture is neutralized with hydrochloric acid, the resulting solid is filtered off, washed with water, dried and mp169-1.
71 ° C N- [4- [4- (4- (2,4-diamino-6-hydroxypyrimidin-5-yl) butyl] benzoyl] -L
Obtain glutamic acid; 1R (KBr) ν _max 3340,3200,2920,
2860, 1600, 1380, 1170 and 755 cm ^-1 ; ¹ H NMR (dTFA) δ 7.3
0 (d, J = 8.7 Hz, 2H, Ar), 6.90 (d, J = 8.7 Hz, 2H, Ar),
4.56-4.61 (m, 1H), 2.26-2.38 (m, 4H, 2 ° aliphatic), 1.
93-2.14 (m, 4H, 2 ° aliphatic), 1.25-1.34 (m, 2H, 2 ° aliphatic), 1.05-1.17 (m, 2H, 2 ° aliphatic). N- [4- [3- (2,4-diamino-6-hydroxypyrimidin-5-yl) propyl] benzoyl] -L-
Glutamic acid, N- [4- [5- (2,4-diamino-6
-Hydroxypyrimidin-5-yl) pentyl] benzoyl] -L-glutamic acid and N- [10- (2,4-diamino-6-hydroxypyrimidin-5-yl) decanoyl] -L-glutamic acid were similarly prepared. It

Example 7 The starting materials used in Example 1 are known or can be prepared by known procedures. The following procedure is representative of such manufacturing.

A. Palladium chloride 0.082 g (0.005 eq) in diethylamine stirred under nitrogen, triphenylphosphine 0.2
44 g (0.01 equivalent) and methyl 4-bromobenzoate 20.00 g
(1.0 equiv.) To the mixture are added 0.178 g (0.01 equiv.) Copper (I) iodide and 6.52 g (1.0 equiv.) 3-butyn-1-ol. The reaction mixture is stirred under nitrogen at room temperature (about 25 ° C) for 18 hours. Jetylamine is then removed under reduced pressure, water is added and the mixture is extracted with benzene. The benzene extract is filtered through silica to remove metal residues and the filtrate is concentrated under reduced pressure to give methyl 4- (4-hydroxybut-1-yn-1-yl) benzoate. Recrystallization from a mixture of benzene and hexane gives pure material with mP 95.5-96.0 ° C in 75.8% yield; 1R (KBr) ν _max 3310,2955,1718,1604,1
433,1275,1177,1108,1040,599.852 and 769 cm ^-1 ; ¹ H NMR
(CDCI ₃ ) δ7.98 (d, J = 8.3 Hz, 2H, Ar), 7.49 (d, J = 8.
3 Hz, 2H, Ar), 3.93 (s, 3H-CH 3) 3.87 (m, 2H, -C H 2 O
H), 2.74 (t, J = 6.2 Hz, 2H, −y1−CH ₂ −), 1.88 (m, 1H,
-OH). Calcd _{(C 12 H 12 O 3)} : C 70.57; H 5.92 Found: C 70.36; instead of H 5.68 3- butyn-1-ol 4-pentyn-1-
By using all, mP68.5-
4- (5-hydroxypent-1-yne-1- at 69.5 ° C
Ile) get methyl benzoate: 1R (KBr) ν _max 3360,295
5,2855,2220,1720,1604,1431,1405,1307,1272,1193,117
2,1112,1063,1017,963,904,859,769 and screw 696 cm ^-1 ; ¹ H
NMR (CDCl ₃ ) δ 7.96 (d, J = 8.3 Hz, 2H, Ar), 7.45 (d, J
= 8.3 Hz, 2H, Ar) , 3.92 (s, 3H, -CH 3), 3.80-3.87 (m,
_{2H, -C H 2 OH),} 2.58 (t, J = 7.0 Hz, 2H, -yl-CH 2 -),
1.63 (bs, 1H, -OH). Calcd _{(C 13 H 14 O 3)} : C 71.54; H 6.47 Found: C 71.26; H 6.38 B. ethanol 200ml of 4 (4-hydroxybutyryl-1-
In-1-yl) 5% in a mixture of 2.55 g of methyl benzoate
50 psi hydrogen in the presence of 0.26 g (10 wt% equivalent) of palladium on charcoal
Hydrogenate (about 3.5 kg / cm ² ) for 12 hours. The reaction mixture is filtered through a silica gel pad, which is washed with ethanol and concentrated to give methyl 4- (4-hydroxybutyl) benzoate as an oil; IR (film) v _max 3390,2
965,2920,2850,1705,1605,1568,15201500,1410,1387,13
62,1308,1286,1250,1160,1055,1013,843,755 and 695 cm
^-1 ; ¹ H NMR (CDCl ₃ ) δ7.95 (d, J = 8.1 Hz, 2H, Ar), 7.25
(D, J = 8.1 Hz, 2H, Ar), 3.89 (s, 3H, -CH 3), 3.65 (t, J
= 6.3 Hz, 2H, -C H 2 OH), 2,69 (t, J = 7.5 Hz, 2H, benzyl) 1.66, (m, 4H, 2 ° aliphatic). Analytical calculation (C ₁₂ H ₁₆ O ₃ ): C 69.21; H 7.74 Found: C 68.97; H 7.92 Methyl 4- (5-hydroxypentyl) benzoate is similarly prepared; 1R (film) ν _max 3380 , 2905,2835,1
700,1598,1562,1424,1405,1300,1266,1168,1097,1058,1
036,1010,953,834,747 and 692 cm ¹ ; ¹ H NMR (CDCl ₃ ) δ 7.
94 (d, J = 8.1 Hz, 2H, Ar), 7.23 (d, J = 8.1 Hz, 2H, Ar),
_{3.89 (s, 3H, -CH 3} ), 3.62 (t, J = 6.5 Hz, 2H, -C H 2 O
H), 2.66 (t, J = 7.7 Hz, 2H, benzyl), 1.86 (bs, 1H, O
H), 1.53-1.71 (m, 4H, 2 ° aliphatic), 1.35-1.45 (m, 2H,
2 ° aliphatic). Analytical calculation (C ₁₃ H ₁₈ O ₃ ): C 70.25; H 8.16 Found: C 70.05; H 8.17

 ─────────────────────────────────────────────────── ─── Continued Front Page (72) Inventor Philip, M, Harrington, Michigan, USA, Plainwell Wedgwood, Drive, 1047 (72) Inventor Chuan, Sea Indiana, Indianapolis, East, Scarsdale, Drive, 8521

Claims (13)

(57) [Claims] 1. A compound of the following formula: [Wherein each of X and Y independently of the other is hydroxy or amino; Z is unsubstituted or substituted with 1 to 4 chlorine or fluorine atoms, 1,4-phenylene; cyclohexa-1, 4-diyl;
Or a linear or branched alkylene group having 2 to 5 carbon atoms; n has a value of 2 to 6; and the carbon atom arrangement shown by * is L] and a pharmaceutically acceptable salt thereof. . 2. The compound according to claim 1, wherein Y is amino and X is hydroxy. 3. The compound according to claim 1, wherein both X and Y are hydroxy. 4. The method of claim 1, wherein both X and Y are amino.
The compound according to the above. 5. Z is 1,4-phenylene, cyclohexa-1,4
-The compound according to claim 1, which is diyl or tetramethylene. 6. The compound according to claim 2, wherein Z is 1,4-phenylene. 7. Z is cyclohexa-1,4-diyl,
A compound according to claim 2. 8. The compound according to claim 4, wherein Z is tetramethylene. 9. — (C _n H _2n ) — is tetramethylene.
A compound according to claim 1. 10. A pharmaceutical composition for combating tumor growth in a mammal, for combating said growth when administered to a mammal in a single or multiple dose therapy with a pharmaceutically acceptable carrier. A composition comprising an effective amount of the compound of claim 1. 11. The following formula: The compound according to claim 1, having the formula: wherein n has a value of 3 to 5 and the carbon atom arrangement indicated by * is L. 12. N- [4- [4- (2,4-diamino-6)
The compound of claim 11, which is -hydroxypyrimidin-5-yl) butyl] benzoyl] -L-glutamic acid. 13. A pharmaceutical composition for combating tumor growth in a mammal for combating said growth when administered to a mammal in a single or multiple dose therapy with a pharmaceutically acceptable carrier. A composition comprising an effective amount of the compound of claim 11.